Fidelity of RNA polymerase II transcription: Role of Rpb9 [corrected] in error detection and proofreading. 2013

Kevin Knippa, and David O Peterson
Department of Biochemistry and Biophysics, Texas A&M University , College Station, Texas 77843-2128, United States.

The role of the small RNA polymerase II subunit Rpb9 in transcriptional proofreading was assessed in vitro. Transcription elongation complexes in which the 3' end of the RNA is not complementary to the DNA template have a dramatically reduced rate of elongation, which provides a fidelity checkpoint at which the error can be removed. The efficiency of such proofreading depends on competing rates of error propagation (extending the RNA chain without removing the error) and error excision, a process that is facilitated by TFIIS. In the absence of Rpb9, the rate of error propagation is increased by 2- to 3-fold in numerous sequence contexts, compromising the efficiency of proofreading. In addition, the rate and extent of TFIIS-mediated error excision is also significantly compromised in the absence of Rpb9. In at least some sequence contexts, Rpb9 appears to enhance TFIIS-mediated error excision by facilitating efficient formation of a conformation necessary for RNA cleavage. If a transcription error is propagated by addition of a nucleotide to the mismatched 3' end, then the rate of further elongation increases but remains much slower than that of a complex with a fully base-paired RNA, which provides a second potential fidelity checkpoint. The absence of Rpb9 also affects both error propagation and TFIIS-mediated error excision at this potential checkpoint in a manner that compromises transcriptional fidelity. In contrast, no effects of Rpb9 on NTP selectivity were observed.

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D012319 RNA Polymerase II A DNA-dependent RNA polymerase present in bacterial, plant, and animal cells. It functions in the nucleoplasmic structure and transcribes DNA into RNA. It has different requirements for cations and salt than RNA polymerase I and is strongly inhibited by alpha-amanitin. EC 2.7.7.6. DNA-Dependent RNA Polymerase II,RNA Pol II,RNA Polymerase B,DNA Dependent RNA Polymerase II
D012441 Saccharomyces cerevisiae A species of the genus SACCHAROMYCES, family Saccharomycetaceae, order Saccharomycetales, known as "baker's" or "brewer's" yeast. The dried form is used as a dietary supplement. Baker's Yeast,Brewer's Yeast,Candida robusta,S. cerevisiae,Saccharomyces capensis,Saccharomyces italicus,Saccharomyces oviformis,Saccharomyces uvarum var. melibiosus,Yeast, Baker's,Yeast, Brewer's,Baker Yeast,S cerevisiae,Baker's Yeasts,Yeast, Baker
D014158 Transcription, Genetic The biosynthesis of RNA carried out on a template of DNA. The biosynthesis of DNA from an RNA template is called REVERSE TRANSCRIPTION. Genetic Transcription
D021122 Protein Subunits Single chains of amino acids that are the units of multimeric PROTEINS. Multimeric proteins can be composed of identical or non-identical subunits. One or more monomeric subunits may compose a protomer which itself is a subunit structure of a larger assembly. Protomers,Protein Subunit,Protomer,Subunit, Protein,Subunits, Protein
D029701 Saccharomyces cerevisiae Proteins Proteins obtained from the species SACCHAROMYCES CEREVISIAE. The function of specific proteins from this organism are the subject of intense scientific interest and have been used to derive basic understanding of the functioning similar proteins in higher eukaryotes. Baker's Yeast Proteins,S cerevisiae Proteins
D035602 Transcriptional Elongation Factors Transcription factors whose primary function is to regulate the rate in which RNA is transcribed. Transcriptional Elongation Inhibitory Factors,Elongation Factors, Transcriptional

Related Publications

Kevin Knippa, and David O Peterson
July 2009, The Journal of biological chemistry,
Kevin Knippa, and David O Peterson
May 1998, Cell,
Kevin Knippa, and David O Peterson
December 2009, Current opinion in structural biology,
Kevin Knippa, and David O Peterson
November 2000, The Journal of biological chemistry,
Kevin Knippa, and David O Peterson
February 2006, Proceedings of the National Academy of Sciences of the United States of America,
Kevin Knippa, and David O Peterson
November 2022, EMBO reports,
Kevin Knippa, and David O Peterson
June 2007, Proceedings of the National Academy of Sciences of the United States of America,
Kevin Knippa, and David O Peterson
January 2000, The Journal of biological chemistry,
Kevin Knippa, and David O Peterson
November 1996, Proceedings of the National Academy of Sciences of the United States of America,
Kevin Knippa, and David O Peterson
December 2003, The Journal of biological chemistry,
Copied contents to your clipboard!